[Histonet] rhodamine auto-fluor. problem

[Glenn Smith]

In a message dated 10/24/2003 3:01:17 PM US Mountain Standard Time,
Bonnie.P.Whitaker <@t> uth.tmc.edu writes:

> I have a question for you guys: A researcher here in OB/GYN is doing
rhodamine-labeled fluorescent work (he didn't say what antibody) on mouse
fallopian tube... he thought he was having a staining problem,  but has
determined that his tissue is auto-fluorescing. The tissue is frozen in OCT
and fixed in acetone/methanol. What can he do to quench this? <

A couple more thoughts Bonnie... 

Rhodamine is excited with 488nm (blue light). I am assuming the system used
in this case employs such a light source. Most organic material fluoresce
under 488nm excitation. The best option is to use a narrow bandpass filter
(20-40nm bandwidth centered about the peak emission wavelength of the dye).
This will not completely eliminate autofluorescence but it will greatly
reduce it.  Another possibility is to image the tissue before and after
staining to see the difference. Then could subtract one image from the
other...

Glenn Smith, P.Eng.
519.886.9013 x38
gsmith <@t> confocal.com
 
Biomedical Photometrics Inc.
A12-550 Parkside Dr.
Waterloo, ON N2L 5V4
 
Widefield High Resolution Imaging Instruments & Software
Please visit www.confocal.com <http://www.confocal.com/>  for more
information.
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